Induction of mesenchymal stem cell differentiation and cartilage formation by cross-linker-free collagen microspheres

• Published in: European cells & materials Vol. 28; pp. 82 - 97
• Main Authors: Mathieu, M, Vigier, S, Labour, M N, Jorgensen, C, Belamie, E, Noël, D
• Format: Journal Article
• Language: English
• Published: Switzerland AO Research Institute Davos 02-09-2014; Forum Multimedia Publishing LLC
• Subjects: Animals; Biomaterial; Cartilage - cytology; Cartilage - physiology; cartilage engineering; Cell Differentiation; Cells, Cultured; Chondrogenesis; collagen; Collagen Type I - chemistry; Collagen Type I - pharmacology; Cross-Linking Reagents - toxicity; Humans; injectable; Life Sciences; mesenchymal stem cell; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - drug effects; Mice; Mice, SCID; Microspheres; Regeneration; safety; self-assembly; Tissue Scaffolds - chemistry; Transforming Growth Factor beta - pharmacology; transforming growth factor-β; mesenchymal stem cell; self-assembly; injectable; microspheres; transforming growth factor-β; safety; cartilage engineering; collagen; Biomaterial; chondrogenesis
• ISSN: 1473-2262; 1473-2262
• DOI: 10.22203/ecm.v028a07

Because of poor self-healing ability, joint cartilage can undergo irreversible degradation in the course of various diseases or after injury. A promising approach for cartilage engineering consists of using of mesenchymal stem cells (MSC) and a differentiation factor combined with an injectable carrier biomaterial. We describe here a novel synthesis route for native collagen microspheres that does not involve the use of potentially toxic crosslinking agents. An emulsion was formed between a type I collagen solution and perfluorinated oil, stabilised by a biocompatible triblock perfluorinated copolymer surfactant. Spherical microparticles of fibrillar collagen were formed through a sol-gel transition induced by ammonia vapours. Electron microscopy observations showed that these self-cross-linked microspheres were constituted by a gel of striated collagen fibrils. Microspheres that were loaded with transforming growth factor beta (TGF-β)3 progressively released this differentiation factor over a four weeks period. Human MSC rapidly adhered to TGF-β3-loaded microspheres and, after 21 d of culture, exhibited typical chondrocyte morphology and produced an uncalcified matrix made of the predominant cartilage components, aggrecan and type II collagen, but devoid of the hypertrophic marker type X collagen. Subcutaneous co-injection of MSC and TGF-β3-loaded microspheres in mice consistently led to the formation of a cartilage-like tissue, which was however hypertrophic, calcified and vascularised. In conclusion, we developed cross-linker free collagen microspheres that allowed chondrogenic differentiation of MSC in vitro and in vivo.